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Laboratory of Molecular and Cellular Physiology  

LabPerego

 

Team leader
Dr.ssa Perego Carla carla.perego@unimi.it                                   

Team
Dr.ssa Eliana Di Cairano, Postdoctoral fellow eliana.dicairano@unimi.it
Dr.ssa Stefania Moretti, PhD Student stefania.moretti@unimi.it
Federica Daniele, Undergraduate fellow federica.danieli@studenti.unimi.it

 

The problem:

The ability of cells to perceive and correctly respond to their microenvironment is the basis of organ development and function. We are interested i) in characterizing the molecular mechanisms of intercellular interactions among neuronal cells in the nervous system, and endocrine cells in islet of Langerhans; and ii) in evaluating the dynamics of cellular responses. Our ultimate goal is to apply the information obtained through these studies toward the development of novel approaches for the treatment of nervous system pathologies and metabolic diseases.

Our current projects include:

1) Dynamic behavior of neurotransmitter transportersin the nervous system and peripheral organs: transporters are emerging as key players in the control of signalling and homeostasis in the nervous system and in the peripheral organs. We are interested in understandinghow these transporters work as molecules, what tasks they fulfil in cell physiology, how they are dynamically regulated, and whether their activity is modified in neurodegenerative diseases.

2) Molecular mechanisms of synaptic transmission in the central nervous system: we are interested in understanding the role of some synaptic proteins in vesicular trafficking dynamics in health and disease.

3) Islets of Langerhans: Pancreatic β-cells play a key role in glucose homeostasis by releasing the glucose-lowering hormone insulin and their death/abnormal activity is implicated in diabetes. Our research is focused in identifying the signals controlling their proliferation, differentiation, activity in physiological conditions and in understanding how these signaling pathways may change in pathological states.

 

The techniques

To achieve these goals, we use cellular and small animal models and we apply state-of-the-art techniques, including:

  • Live cell Imaging, we use GFP-tagged proteins or organic dyes and we track signaling dynamics by means of: 

- Ion imaging to follow changes in ion concentrations induced by activation of receptors, channels, transporters

- Total Internal Reflection Fluorescence Microscopy (TIRFM) to study the events taking place at/or immediately; beneath the plasma membrane, i.e. protein/vesicle exo- or endo-cytosis, protein clustering;  

- We are setting up Fluorescence Resonance Energy Transfer (FRET) to follow the dynamics of protein interactions;

- Confocal microscopy to follow the distribution and the trafficking of proteins in cells and small animals.

  • Molecular biology techniques: RT-PCR, Site-directed mutagenesis, functional expression cloning, Northern blotting.
  • Biochemistry technique: Western blots; Immunoprecipitation; Generation, expression and purification of Recombinant proteins (GST/His/GTRAP-tagged); Affinity chromatography assays; ELISA assays.

 

Selection of Pubblications

1. Perego C, Cairano ES, Ballabio M, Magnaghi V. (2012) Neurosteroid allopregnanolone regulates EAAC1-mediated glutamate uptake and triggers actin changes in Schwann cells. J Cell Physiol.227(4):1740-51.

2. Davalli AM, Perego C, Folli F. (2012) The potential role of glutamate in the current diabetic epidemic. Acta Diabetol. 2012 Jan 5. DOI: 10.1007/s00592-011-0364-z

3. Folli F, Okada T, Perego C, Gunton G, Liew CW, Akiyama M, D’Amico A, Larosa S, Placidi C, Lupi R, Marchetti P, Sesti G, Hellerstein M, Perego L, Kulkarni RN. (2011) Altered Insulin Receptor Signalling and β-Cell Cycle Dynamics in Type 2 Diabetes Mellitus. PLoS One6(11):e28050.  

4. Di Cairano ES, Davalli AM, Perego L, Sala S, Sacchi VF, La Rosa S, Placidi C, Cappella C, Conti P, Centonze VE, Casiraghi F, Bertuzzi F, Folli F, and Perego C. (2011) The glial glutamate transporter 1 (GLT1) is expressed by pancreatic beta-cells and prevents glutamate-induced beta-cell death. J Biol Chem.286 (16):14007-18.

5. D’Amico A, Soragna A, Di Cairano E, Panzeri N, Anzai N, Sacchi VF and Perego C. (2010) The surface density of the glutamate transporter EAAC1 is controlled by interactions with PDZK1 and AP2 adaptor complexes. Traffic11(11):1455-70.

6. Guardado-Mendoza R, Davalli A, Chavez A, Hubbard G, Dick E, Majluf-Cruz A, Tene-Perez C, Goldschmidt L, Hart J, Perego C, Comuzzie A, Tejero M.E, Finzi G, Placidi C, La Rosa S, Capella C, Halff G, Gastaldelli A, DeFronzo R, Folli F. (2009) Islet Amyloidosis, Beta Cell Apoptosis and Alpha Cell Proliferation are Key Determinants of Islets’ of Langerhans Dysfunctional Remodeling and Hyperglycemia. Development of a Novel Predictive Model of Type 2 Diabetes Mellitus in the Baboon. Proc Natl Acad Sci USA106(33):13992-7.

7. Lenardi C, Perego C, Cassina V, Podestà A, D’Amico A, Gualandris D, Vinati S, Fiorentini F, Bongiorno G, Piseri P, Sacchi VF, Milani P. (2006) Adhesion and proliferation of fibroblasts on cluster-assembled nanostructured carbon films: the role of surface morphology. J.  Nanoscience and Nanotechnology. 6(12):3718-30. 

8. Perego C., Vanoni C., Massari S., Longhi R.and Pietrini G. (2000) Mammalian LIN-7 PDZ proteins associate with β-catenin at the cell-cell junctions of epithelia and neurons. EMBO J. 19: 3978-89.

9. Perego C., Vanoni C., Bossi M., Massari S., Basudev H., Longhi R., and Pietrini G. (2000) The GLT-1 and GLAST glutamate transporters are expressed on morphologically distinct astrocytes and regulated by neuronal activity in primary hippocampal cocultures. J. Neurochem.75: 1076-84.

10. Perego C, Vanoni C, Villa A, Longhi R, Kaech S.M, Frohli E, Hajnal  A, Kim S.K and Pietrini G. (1999) PDZ-mediated interactions retain the epithelial GABA transporter on the basolateral surface of polarized epithelial cells. EMBO J. 18: 2384-93.

 

Brevetti

Perego C, Di Cairano ES, Davalli A, Folli F. Methods and compositions for the diagnosis and treatment of diabetes” US Patent.

 
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